Recent Advances in HPLC-MS for Pharmaceutical Analysis

High-performance liquid chromatography-mass spectrometry (HPLC-MS) combines the separation power of LC with the detection specificity of MS. It has become an indispensable analytical tool in the pharmaceutical industry due to its high throughput, sensitivity, and selectivity.

Image Credit: Sodel Vladyslav/

Image Credit: Sodel Vladyslav/

HPLC-MS is utilized throughout the drug development process: it has applications in product characterization, drug discovery, metabolism studies (in vitro and in vivo), and the identification of impurities and degradation products.1 However, the growing demand for drug discovery has challenged the applications of HPLC-MS in pharmaceutical analysis: the industry now requires increased production for the same time and monetary cost. Automated, high-throughput analysis is therefore required. Innovative approaches in HPLC-MS are thus necessary to maintain pace with industry demands.

Pharmaceuticals and Biologics’ is one of the six tracks at Pittcon 2024, where attendees can learn about the latest developments in pharmaceutical analysis from experts in the field. This article highlights some of these groundbreaking innovations, setting the stage for the upcoming event.

Current Developments in HPLC-MS

HPLC-MS interface technologies are continuously evolving, with significant advancements occurring in areas such as automated sample preparation, parallel LC separation, and the enhancement of MS detection efficiency.

Sample preparation is the first step in HPLC-MS: it is required to remove interfering components from a complex matrix. This process is typically laborious and is often considered the bottleneck step in bioanalysis.2 The sample preparation techniques commonly used in the pharmaceutical field are solid-phase extraction, liquid-liquid extraction, and supported liquid extraction. However, approaches to automate these methods are being made to help combat the demand for rapid, high-throughput analysis.

Another way to enhance throughput is by using multiple columns in parallel. With parallel HPLC-MS, a single sample injection is divided into different columns for separation and subsequently recombined for analysis via a single multi-channel absorbance detector.3 This increases the number of samples that can be analyzed while maintaining separation efficiency.

Continuous efforts to enhance the sensitivity of MS detection have also been made, including the improvement of pretreatment procedures, ionization techniques, and mass analyzers.4 The demand for quicker testing, higher throughput, and smaller footprints has also driven the industry to develop small, portable MS systems.

Pittcon 2024: Unveiling the Future of HPLC-MS

At Pittcon 2024, industry experts will provide attendees with valuable insights into existing and up-and-coming trends in HPLC-MS for drug development and discovery. Pittcon also opens the door for networking opportunities with market-leading analytical equipment manufacturers. This year, GenTech Scientific is exhibiting, giving those who come along to the event that chance to witness first-hand some of the most cutting-edge HPLC-MS instrumentation to date.

At Pittcon 2024, Dr. Perry Wang, Research Chemist at the US FDA, will host an organized session titled “High Throughput Drug Analysis by LC-MS and Its Challenges.”5 Leveraging his expertise in HPLC-MS method development and validation for drugs, foods, cosmetics, and dietary supplements, Dr. Wang will provide attendees with a comprehensive perspective on innovative HPLC-MS techniques designed to increase throughput without compromising sensitivity and selectivity.

His recent work explored how multidimensional separation methods—with improved sensitivity, peak capacity, and throughput—can enhance separation performance for complex proteomics samples and enable profiling of low-microgram samples.At Pittocon, Dr. Wang will offer insight into how these advancements are expected to shape pharmaceutical analysis in the coming decades.

Dr. Aihua Liu, Senior Director of Bioanalysis at Resolian, will also present at Pittcon 2024. She has over fifteen years of experience in bioanalysis, with expertise in analytical method development and validation for small molecules, oligonucleotides, antibody-drug conjugates, lipids, Dibenzylidene sorbitol, and more. Her talk, titled “Pros and Cons of Different Chromatographic Technologies in High-Throughput Bioanalysis of Oligonucleotide Therapeutics,”7 will highlight the need for precise and efficient bioanalysis of oligonucleotide therapeutics (ONs).

Bioanalysis of ONs: The Advantages and Disadvantages

ONs consist of short synthetic strands of RNA or DNA. They have recently emerged as potential medical therapeutics due to their ability to modulate gene expression.

Three types of ONs exist—small interference RNA (siRNA), anti-sense oligonucleotides (ASOs), and aptamer RNAs. The initial generations of ONs faced substantial challenges that demanded innovative solutions via chemical modifications and complex delivery systems.

However, the incorporation of these solutions introduced new challenges in ONs bioanalysis, resulting in the deployment of numerous chromatographic technologies—including ion-pairing LC (IP-LC), reverse-phased LC (RP-LC), hydrophilic interaction LC (HILIC), and ion exchange LC (IE-LC).

Dr. Liu and her team have extensive experience developing and validating these techniques for ONs bioanalysis—including IP-LC-MS/MS, HILIC-LC-MS/MS, RP-LC-MS/MS, IE-LC-Fluorescence Detection (IE-LC-FLD), and P-LC-High-Resolution MS (IP-LC-HRMS). In her talk at Pittcon, Dr. Liu will evaluate the use of these chromatographic technologies for the high-throughput analysis of different ONs (including siRNA and ASOs of variable lengths). Real-case studies will also be utilized to highlight the strengths and weaknesses of each technique and provide valuable insights into the high-throughput bioanalysis of ONs.

Find out more at Pittcon 2024

The applications of HPLC-MS in discovering novel drug candidates, studying drug metabolism, and identifying and characterizing impurities and degradation products have risen exponentially.

Undoubtedly, high-throughput analysis by HPLC-MS will continue to play an important role in the pharmaceutical field over the next few decades. However, to keep up with the demand for new chemical entities, innovations in sample preparation and HPLC-MS technologies are required.

This will be addressed at Pittcon 2024, where attendees will gain valuable insights into the current and future trends of HPLC-MS in pharmaceutical analysis.

To find out more about Pittcon and register, please visit the Pittcon website.

References and Further Reading

  1. Beccaria, M., et al. (2020). Current Developments in LC-MS for Pharmaceutical Analysis. Analyst.
  2. Ingle, RG., et al. (2022). Current Developments of Bioanalytical Sample Preparation Techniques in Pharmaceuticals. The Journal of Pharmaceutical Analysis.
  3. Gross, G.M., et al. (2003). Parallel Column Liquid Chromatography with a Single Multi-Wavelength Absorbance Detector for Enhanced Selectivity Using Chemometric Analysis. Analytica Chimica Acta.
  4. Li, C., et al. (2021). Towards Higher Sensitivity of Mass Spectrometry: A Perspective From the Mass Analyzers. Frontiers in Chemistry.
  5. High Throughput Drug Analysis by LC-MS and Its Challenges (no date) Attend networking events and meet the right people with the LabSci by Pittcon event app. Available at: (Accessed: 5 December 2023)
  6. Gang, L., et al. (2023). Multidimensional Separation Methods Can Improve Separation Performance for Extremely Complicated Proteomics Samples and Provide a Powerful Tool for Comprehensive Proteome Profiling of Low-Microgram Samples. Bioanalysis.
  7. Pros and Cons of Different Chromatographic Technologies in High-Throughput Bioanalysis of Oligonucleotide Therapeutics (no date) Attend networking events and meet the right people with the LabSci by Pittcon event app. Available at: (Accessed: 5 December 2023).

About Pittcon

Pittcon is the world’s largest annual premier conference and exposition on laboratory science. Pittcon attracts more than 16,000 attendees from industry, academia and government from over 90 countries worldwide.

Their mission is to sponsor and sustain educational and charitable activities for the advancement and benefit of scientific endeavor.

Pittcon’s target audience is not just “analytical chemists,” but all laboratory scientists — anyone who identifies, quantifies, analyzes or tests the chemical or biological properties of compounds or molecules, or who manages these laboratory scientists.

Having grown beyond its roots in analytical chemistry and spectroscopy, Pittcon has evolved into an event that now also serves a diverse constituency encompassing life sciences, pharmaceutical discovery and QA, food safety, environmental, bioterrorism and other emerging markets.

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Last updated: Dec 5, 2023 at 7:14 AM


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